Alkylamido Compounds and Uses Thereof

ABSTRACT

Disclosed herein are compounds that may be specific to PPAR and/or EGF receptors, and methods of making and using same.

This application is a continuation application of U.S. patentapplication Ser. No. 16/149,524, filed on Oct. 2, 2018, whichapplication is a continuation application of U.S. patent applicationSer. No. 15/337,707, filed on Oct. 28, 2016, which is a continuationapplication of U.S. patent application Ser. No. 14/255,255, filed onApr. 17, 2014, which is a continuation application of U.S. patentapplication Ser No. 13/201,786, filed on Nov. 17, 2011, which is a U.S.national stage application under 35 U.S.C. § 371 of InternationalApplication No. PCT/EP2010/000935, filed on Feb. 16, 2010, which claimspriority to European Application No. 09425056.0, filed on Feb. 16, 2009,U.S. Patent Application No. 61/179,062, filed on May 18, 2009 and U.S.Patent Application No. 61/287,461, filed on Dec. 17, 2009, each of whichis incorporated by reference herein in its entirety.

BACKGROUND

Peroxisome Proliferator Activated Receptors (PPARs) are members of thenuclear hormone receptor super family, which are ligand-activatedtranscription factors regulating gene expression. Certain PPARs playroles in the regulation of cell differentiation, development andmetabolism of higher organisms.

Three types of PPAR has been identified: alpha, expressed in the liver,kidney, heart and other tissues and organs, beta/delta expressed forexample in the brain, and gamma, expressed in three forms: gammal,gamma2, and gamma3. PPARγ receptors have been associated with a numberof disease states including dyslipidemia, hyperlipidemia,hypercholesteremia, atherosclerosis, atherogenesis,hypertriglyceridemia, heart failure, myocardial infarction, vasculardiseases, cardiovascular diseases, hypertension, obesity, inflammation,arthritis, cancer, Alzheimer's disease, skin disorders, respiratorydiseases, ophthalmic disorders, IBDs (irritable bowel disease),ulcerative colitis and Crohn's disease.

Further, treatment of tumor cells with ligands of PPARγ receptors caninduce a decrease in cellular proliferation, cell differentiation andapoptosis, and therefore may be useful in preventing carcinogenesis.Intestinal anti-inflammatory activity may be dependent on binding andsubsequent activation of PPARγ receptors.

In addition, numerous studies have indicated that EGF receptorinhibitors may control proliferation and spread of tumors.

Accordingly, molecules that modulate the activity of PPARs and/or EGFreceptors are useful as therapeutic agents in the treatment of suchdiseases.

SUMMARY

This disclosure is generally directed to compounds which may be specificto PPAR receptors and/or EGF receptors, and their use as, for example,medicinal agents. Also provided are pharmaceutical compositionscomprising at least one disclosed compound, or pharmaceuticallyacceptable salt or N-oxide thereof, and a pharmaceutically acceptablecarrier.

One embodiment provides compounds represented by formula I, andcompositions comprising such compounds:

-   -   wherein X is C₁-C₃alkylene, optionally substituted with one, two        or three substituents selected from halogen or hydroxyl;    -   R₁ is selected from the group consisting of C₁-C₆alkyl,        C₃-C₆cycloalkyl, C₂-C₆alkenyl, and C₂-C₆alkynyl;    -   R₂ is selected from the group consisting of hydrogen and        C₁-C₆alkyl;    -   R₃ is independently selected, for each occurrence from the group        consisting of hydrogen, C₁-C₆alkoxy, C₁-C₆alkyl, cyano,        C₃-C₆cycloalkyl, halogen, hydroxyl, and nitro;    -   R₄ is selected from the group consisting of hydrogen and        C₁-C₆alkyl;    -   R₅ is hydrogen or C₁-C₆alkyl;    -   or pharmaceutically acceptable salts or N-oxides thereof.

Another embodiment provides compounds represented by formula II, andcompositions comprising such compounds:

-   -   wherein R₁ is selected from the group consisting of C₁-C₆alkyl,        C₃-C₆cycloalkyl, C₂-C₆alkenyl, and C₂-C₆alkynyl;    -   R₂ is selected from the group consisting of hydrogen and        C₁-C₆alkyl;    -   R₃ is independently selected, for each occurrence from the group        consisting of hydrogen, C₁-C₆alkoxy, C₁-C₆alkyl, cyano,        C₃-C₆cycloalkyl, halogen, hydroxyl, and nitro;    -   R₅ is C₁-C₆alkyl;        or pharmaceutically acceptable salts or N-oxides thereof.

Also provided herein are methods of treating cancer (e.g. colorectalcancer) such as tumors expressing PPAR receptors and/or EGF receptors,comprising administering to a subject in need thereof a therapeuticallyeffective amount of a compound of formula I or II, or a pharmaceuticallyacceptable salt or N-oxides thereof. Also contemplated herein arecompositions that include a compound represented by formula I or II ande.g., a pharmaceutically acceptable excipient.

Also provided are compounds represented by formulas I and II for use intherapy and/or for the manufacture of a medicament for the treatment ofcancer such as colorectal cancer.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 depicts the % mortality of mice receiving TNBS (trinitrobenzenesulfonic acid) versus mice receiving TNBS and N-acetyl E2 in a murinecolitis model.

FIG. 2 depicts the observed level of colonic lesions in mice receivingTNBS versus mice receiving TNBS and N-acetyl E2 in a murine colitismodel.

FIG. 3 depicts the observed level of MPO activity in mice receiving TNBSversus mice receiving TNBS and N-acetyl E2 in a murine colitis model.

FIG. 4 depicts the observed level of colonic inflammation in micereceiving TNBS versus mice receiving TNBS and N-acetyl E2 in a murinecolitis model.

FIG. 5 depicts effects of a disclosed compound on human keratinocytes.

FIG. 6 depicts inhibition of TNF alpha by H₂O₂ and a disclosed compound.

FIG. 7 depicts inhibition on mRNA expression of IL-6 induced by thepresence of IFN-gamma.

FIG. 8 depicts inhibition of a disclosed compound on the activation ofNF-kB.

FIG. 9 depicts inhibition of a disclosed compound on protein expressionof IL-6 induced by presence of LPS.

FIG. 10 depicts effect of a disclosed compound on human sebocytes.

FIG. 11 depicts inhibitory capacity of a disclosed compound onsebogenesis induced by lipid type stimulus.

FIG. 12A and FIG. 12B depict the results of a fatty acid assay andsqualene analysis of sebogenesis inhibition, respectively.

FIG. 13 depicts treatment with linoleic acid and testosterone withlipidogenic stimulus.

DETAILED DESCRIPTION

The features and other details of the disclosure will now be moreparticularly described. Before further description of the presentinvention, certain terms employed in the specification, examples andappended claims are collected here. These definitions should be read inlight of the remainder of the disclosure and understood as by a personof skill in the art. Unless defined otherwise, all technical andscientific terms used herein have the same meaning as commonlyunderstood by a person of ordinary skill in the art.

Definitions

“Treating” includes any effect, e.g., lessening, reducing, modulating,or eliminating, that results in the improvement of the condition,disease, disorder and the like.

The term “alkenyl” as used herein refers to an unsaturated straight orbranched hydrocarbon having at least one carbon-carbon double bond, suchas a straight or branched group of 2-12, 2-10, or 2-6 carbon atoms,referred to herein as C₂-C₁₂alkenyl, C₂-C₁₀alkenyl, and C₂-C₆alkenyl,respectively. Exemplary alkenyl groups include, but are not limited to,vinyl, allyl, butenyl, pentenyl, hexenyl, butadienyl, pentadienyl,hexadienyl, 2-ethylhexenyl, 2-propyl-2-butenyl,4-(2-methyl-3-butene)-pentenyl, etc.

The term “alkoxy” as used herein refers to an alkyl group attached to anoxygen (—O-alkyl-). Exemplary alkoxy groups include, but are not limitedto, groups with an alkyl, alkenyl or alkynyl group of 1-12, 1-8, or 1-6carbon atoms, referred to herein as C₁-C₁₂alkoxy, C₁-C₈alkoxy, andC₁-C₆alkoxy, respectively. Exemplary alkoxy groups include, but are notlimited to methoxy, ethoxy, etc. Similarly, exemplary “alkenoxy” groupsinclude, but are not limited to vinyloxy, allyloxy, butenoxy, etc.

The term “alkyl” as used herein refers to a saturated straight orbranched hydrocarbon, such as a straight or branched group of 1-12,1-10, or 1-6 carbon atoms, referred to herein as C₁-C₁₂alkyl,C₁-C₁₀alkyl, and C₁-C₆alkyl, respectively. Exemplary alkyl groupsinclude, but are not limited to, methyl, ethyl, propyl, isopropyl,2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl,3-methyl-1-butyl, 2-methyl-3-butyl, 2,2-dimethyl-1-propyl,2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl,2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl,2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, butyl,isobutyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, heptyl, octyl,etc. In certain embodiments, alkyl refers to C₁-C₆ alkyl. In certainembodiments, cycloalkyl refers to C₃-C₆cycloalkyl.

Alkyl, alkenyl and alkynyl groups can, in some embodiments, beoptionally be substituted with or interrupted by at least one groupselected from alkanoyl, alkoxy, alkyl, alkenyl, alkynyl, amido, amidino,amino, aryl, arylalkyl, azido, carbamate, carbonate, carboxy, cyano,cycloalkyl, ester, ether, formyl, halogen, haloalkyl, heteroaryl,heterocyclyl, hydroxyl, imino, ketone, nitro, phosphate, phosphonato,phosphinato, sulfate, sulfide, sulfonamido, sulfonyl and thiocarbonyl.

The term “alkynyl” as used herein refers to an unsaturated straight orbranched hydrocarbon having at least one carbon-carbon triple bond, suchas a straight or branched group of 2-12, 2-8, or 2-6 carbon atoms,referred to herein as C₂-C₁₂alkynyl, C₂-C₈alkynyl, and C₂-C₆alkynyl,respectively. Exemplary alkynyl groups include, but are not limited to,ethynyl, propynyl, butynyl, pentynyl, hexynyl, methylpropynyl,4-methyl-1-butynyl, 4-propyl-2-pentynyl, and 4-butyl-2-hexynyl, etc.

The term “amide” or “amido” as used herein refers to a radical of theform —R_(a)C(O)N(R_(b))—, —R_(a)C(O)N(R_(b))R_(c)—, or —C(O)NR_(b)R_(c),wherein R_(a), R_(b) and R_(c) are each independently selected fromalkoxy, alkyl, alkenyl, alkynyl, amide, amino, aryl, arylalkyl,carbamate, cycloalkyl, ester, ether, formyl, halogen, haloalkyl,heteroaryl, heterocyclyl, hydrogen, hydroxyl, ketone, and nitro. Theamide can be attached to another group through the carbon, the nitrogen,R_(b), R_(c), or R_(a). The amide also may be cyclic, for example R_(b)and R_(c), R_(a) and R_(b), or R_(a) and R_(c) may be joined to form a3- to 12-membered ring, such as a 3- to 10-membered ring or a 5- to6-membered ring. The term “carboxamido” refers to the structure—C(O)NR_(b)R_(c).

The term “amidino” as used herein refers to a radical of the form—C(═NR)NR′R″ where R, R′, and R″ can each independently be selected fromalkyl, alkenyl, alkynyl, amide, aryl, arylalkyl, cyano, cycloalkyl,haloalkyl, heteroaryl, heterocyclyl, hydroxyl, ketone and nitro.

The term “amine” or “amino” as used herein refers to a radical of theform —NR_(d)R_(e), —N(R_(d))R_(e)—, or —R_(c)N(R_(d))R_(f)— where R_(d),R_(e), and R_(f) are independently selected from alkoxy, alkyl, alkenyl,alkynyl, amide, amino, aryl, arylalkyl, carbamate, cycloalkyl, ester,ether, formyl, halogen, haloalkyl, heteroaryl, heterocyclyl, hydrogen,hydroxyl, ketone, and nitro. The amino can be attached to the parentmolecular group through the nitrogen, R_(d), R_(e) or R_(f). The aminoalso may be cyclic, for example any two of Rd, Re or Rf may be joinedtogether or with the N to form a 3- to 12-membered ring, e.g.,morpholino or piperidinyl. The term amino also includes thecorresponding quaternary ammonium salt of any amino group, e.g.,—[N(Rd)(Re)(Rf)]+. Exemplary amino groups include aminoalkyl groups,wherein at least one of R_(d), R_(e), or R_(f) is an alkyl group.

The term “aryl” as used herein refers to refers to a mono-, bi-, orother multi-carbocyclic, aromatic ring system. In certain embodiments,aryl refers to a monocyclic and/or bicyclic, 5 to 6 membered ring. Thearomatic ring may be substituted at one or more ring positions withsubstituents selected from alkanoyl, alkoxy, alkyl, alkenyl, alkynyl,amido, amidino, amino, aryl, arylalkyl, azido, carbamate, carbonate,carboxy, cyano, cycloalkyl, ester, ether, formyl, halogen, haloalkyl,heteroaryl, heterocyclyl, hydroxyl, imino, ketone, nitro, phosphate,phosphonato, phosphinato, sulfate, sulfide, sulfonamido, sulfonyl andthiocarbonyl. The term “aryl” also includes polycyclic ring systemshaving two or more cyclic rings in which two or more carbons are commonto two adjoining rings (the rings are “fused rings”) wherein at leastone of the rings is aromatic, e.g., the other cyclic rings may becycloalkyls, cycloalkenyls, cycloalkynyls, and/or aryls. Exemplary arylgroups include, but are not limited to, phenyl, tolyl, anthracenyl,fluorenyl, indenyl, azulenyl, and naphthyl, as well as benzo-fusedcarbocyclic moieties such as 5,6,7,8-tetrahydronaphthyl.

The term “arylalkyl” as used herein refers to an aryl group having atleast one alkyl substituent, e.g. -aryl-alkyl-. Exemplary arylalkylgroups include, but are not limited to, arylalkyls having a monocyclicaromatic ring system, wherein the ring comprises 6 carbon atoms. Forexample, “phenylalkyl” includes phenylC4alkyl, benzyl, 1-phenylethyl,2-phenylethyl, etc.

The term “carbonyl” as used herein refers to the radical —C(O)—.

The term “carboxamido” as used herein refers to the radical —C(O)NRR′,where R and R′ may be the same or different. R and R′ may be selectedfrom, for example, alkyl, aryl, arylalkyl, cycloalkyl, formyl,haloalkyl, heteroaryl and heterocyclyl.

The term “carboxy” as used herein refers to the radical —COOH or itscorresponding salts, e.g. —COONa, etc.

The term “cyano” as used herein refers to the radical —CN.

The term “cycloalkoxy” as used herein refers to a cycloalkyl groupattached to an oxygen.

The term “cycloalkyl” as used herein refers to a monovalent saturated orunsaturated cyclic, bicyclic, or bridged bicyclic hydrocarbon group of3-12, 3-8, 4-8, or 4-6 carbons, referred to herein, e.g., as“C₄₋₈cycloalkyl,” derived from a cycloalkane. Exemplary cycloalkylgroups include, but are not limited to, cyclohexanes, cyclohexenes,cyclopentanes, cyclopentenes, cyclobutanes and cyclopropanes. Cycloalkylgroups may be substituted with alkanoyl, alkoxy, alkyl, alkenyl,alkynyl, amido, amidino, amino, aryl, arylalkyl, azido, carbamate,carbonate, carboxy, cyano, cycloalkyl, ester, ether, formyl, halogen,haloalkyl, heteroaryl, heterocyclyl, hydroxyl, imino, ketone, nitro,phosphate, phosphonato, phosphinato, sulfate, sulfide, sulfonamido,sulfonyl and thiocarbonyl. Cycloalkyl groups can be fused to othercycloalkyl, aryl, or heterocyclyl groups. In certain embodiments,cycloalkyl refers to C₃-C₆ alkyl.

The terms “halo” or “halogen” or “Hal” as used herein refer to F, Cl,Br, or I.

The term “haloalkyl” as used herein refers to an alkyl group substitutedwith one or more halogen atoms.

The term “nitro” as used herein refers to the radical —NO₂.

The term “phenyl” as used herein refers to a 6-membered carbocyclicaromatic ring. The phenyl group can also be fused to a cyclohexane orcyclopentane ring. Phenyl can be substituted with one or moresubstituents including alkanoyl, alkoxy, alkyl, alkenyl, alkynyl, amido,amidino, amino, aryl, arylalkyl, azido, carbamate, carbonate, carboxy,cyano, cycloalkyl, ester, ether, formyl, halogen, haloalkyl, heteroaryl,heterocyclyl, hydroxyl, imino, ketone, nitro, phosphate, phosphonato,phosphinato, sulfate, sulfide, sulfonamido, sulfonyl and thiocarbonyl.

The term “phosphate” as used herein refers to the radical—OP(O)(OR_(aa))₂ or its anions. The term “phosphanate” refers to theradical —P(O)(OR_(aa))₂ or its anions. The term “phosphinate” refers tothe radical —PR_(aa)(O)(OR_(aa)) or its anion, where each R_(aa) can beselected from, for example, alkyl, alkenyl, alkynyl, aryl, arylalkyl,cycloalkyl, hydrogen, haloalkyl, heteroaryl, and heterocyclyl.

The term “pharmaceutically acceptable carrier” or “pharmaceuticallyacceptable excipient” as used herein refers to any and all solvents,dispersion media, coatings, isotonic and absorption delaying agents, andthe like, that are compatible with pharmaceutical administration. Theuse of such media and agents for pharmaceutically active substances iswell known in the art. The compositions may also contain other activecompounds providing supplemental, additional, or enhanced therapeuticfunctions.

The term “pharmaceutical composition” as used herein refers to acomposition comprising at least one compound as disclosed hereinformulated together with one or more pharmaceutically acceptablecarriers.

“Individual,” “patient,” or “subject” are used interchangeably andinclude to any animal, including mammals, preferably mice, rats, otherrodents, rabbits, dogs, cats, swine, cattle, sheep, horses, or primates,and most preferably humans. The compounds of the invention can beadministered to a mammal, such as a human, but can also be other mammalssuch as an animal in need of veterinary treatment, e.g., domesticanimals (e.g., dogs, cats, and the like), farm animals (e.g., cows,sheep, pigs, horses, and the like) and laboratory animals (e.g., rats,mice, guinea pigs, and the like). The mammal treated in the methods ofthe invention is desirably a mammal in whom modulation of PPAR and/orEGF receptors is desired. “Modulation” includes antagonism (e.g.,inhibition), agonism, partial antagonism and/or partial agonism.

In the present specification, the term “therapeutically effectiveamount” means the amount of the subject compound that will elicit thebiological or medical response of a tissue, system, animal or human thatis being sought by the researcher, veterinarian, medical doctor or otherclinician. The compounds of the invention are administered intherapeutically effective amounts to treat a disease. Alternatively, atherapeutically effective amount of a compound is the quantity requiredto achieve a desired therapeutic and/or prophylactic effect, such as anamount which results in the prevention of or a decrease in the symptomsassociated with a disease associated with PPAR and/or EGF receptors.

The term “pharmaceutically acceptable salt(s)” as used herein refers tosalts of acidic or basic groups that may be present in compounds used inthe present compositions. Compounds included in the present compositionsthat are basic in nature are capable of forming a wide variety of saltswith various inorganic and organic acids. The acids that may be used toprepare pharmaceutically acceptable acid addition salts of such basiccompounds are those that form non-toxic acid addition salts, i.e., saltscontaining pharmacologically acceptable anions, including but notlimited to malate, oxalate, chloride, bromide, iodide, nitrate, sulfate,bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate,salicylate, citrate, tartrate, oleate, tannate, pantothenate,bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate,gluconate, glucuronate, saccharate, formate, benzoate, glutamate,methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonateand pamoate (i.e., 1,1′-methylene-bis-(2-hydroxy-3-naphthoate)) salts.Compounds included in the present compositions that include an aminomoiety may form pharmaceutically acceptable salts with various aminoacids, in addition to the acids mentioned above. Compounds included inthe present compositions that are acidic in nature are capable offorming base salts with various pharmacologically acceptable cations.Examples of such salts include alkali metal or alkaline earth metalsalts and, particularly, calcium, magnesium, sodium, lithium, zinc,potassium, and iron salts.

The compounds of the disclosure may contain one or more chiral centersand/or double bonds and, therefore, exist as stereoisomers, such asgeometric isomers, enantiomers or diastereomers. The term“stereoisomers” when used herein consist of all geometric isomers,enantiomers or diastereomers. These compounds may be designated by thesymbols “R” or “S,” depending on the configuration of substituentsaround the stereogenic carbon atom. The present invention encompassesvarious stereoisomers of these compounds and mixtures thereof.Stereoisomers include enantiomers and diastereomers. Mixtures ofenantiomers or diastereomers may be designated “(±)” in nomenclature,but the skilled artisan will recognize that a structure may denote achiral center implicitly.

Individual stereoisomers of compounds of the present invention can beprepared synthetically from commercially available starting materialsthat contain asymmetric or stereogenic centers, or by preparation ofracemic mixtures followed by resolution methods well known to those ofordinary skill in the art. These methods of resolution are exemplifiedby (1) attachment of a mixture of enantiomers to a chiral auxiliary,separation of the resulting mixture of diastereomers byrecrystallization or chromatography and liberation of the optically pureproduct from the auxiliary, (2) salt formation employing an opticallyactive resolving agent, or (3) direct separation of the mixture ofoptical enantiomers on chiral chromatographic columns. Stereoisomericmixtures can also be resolved into their component stereoisomers by wellknown methods, such as chiral-phase gas chromatography, chiral-phasehigh performance liquid chromatography, crystallizing the compound as achiral salt complex, or crystallizing the compound in a chiral solvent.Stereoisomers can also be obtained from stereomerically-pureintermediates, reagents, and catalysts by well known asymmetricsynthetic methods.

Geometric isomers can also exist in the compounds of the presentinvention. The symbol

denotes a bond that may be a single, double or triple bond as describedherein. The present invention encompasses the various geometric isomersand mixtures thereof resulting from the arrangement of substituentsaround a carbon-carbon double bond or arrangement of substituents arounda carbocyclic ring. Substituents around a carbon-carbon double bond aredesignated as being in the “Z” or “E” configuration wherein the terms“Z” and “E” are used in accordance with IUPAC standards. Unlessotherwise specified, structures depicting double bonds encompass boththe “E” and “Z” isomers.

Substituents around a carbon-carbon double bond alternatively can bereferred to as “cis” or “trans,” where “cis” represents substituents onthe same side of the double bond and “trans” represents substituents onopposite sides of the double bond. The arrangement of substituentsaround a carbocyclic ring are designated as “cis” or “trans.” The term“cis” represents substituents on the same side of the plane of the ringand the term “trans” represents substituents on opposite sides of theplane of the ring. Mixtures of compounds wherein the substituents aredisposed on both the same and opposite sides of plane of the ring aredesignated “cis/trans.”

The compounds disclosed herein can exist in solvated as well asunsolvated forms with pharmaceutically acceptable solvents such aswater, ethanol, and the like, and it is intended that the inventionembrace both solvated and unsolvated forms. In one embodiment, thecompound is amorphous. In one embodiment, the compound is a polymorph.In another embodiment, the compound is in a crystalline form.

The invention also embraces isotopically labeled compounds of theinvention which are identical to those recited herein, except that oneor more atoms are replaced by an atom having an atomic mass or massnumber different from the atomic mass or mass number usually found innature. Examples of isotopes that can be incorporated into compounds ofthe invention include isotopes of hydrogen, carbon, nitrogen, oxygen,phosphorus, fluorine and chlorine, such as ²H, ³H, ¹³C, ¹⁴ _(C,) ¹⁵_(N,) ¹⁸O, ¹⁷O, ³¹P, ³²P, ³⁵S, ¹⁸F, and ³⁶Cl, respectively.

Certain isotopically-labeled disclosed compounds (e.g., those labeledwith ³H and ¹⁴C) are useful in compound and/or substrate tissuedistribution assays. Tritiated (i.e., ³H) and carbon-14 (i.e., ¹⁴C)isotopes are particularly preferred for their ease of preparation anddetectability. Further, substitution with heavier isotopes such asdeuterium (i.e., ²H) may afford certain therapeutic advantages resultingfrom greater metabolic stability (e.g., increased in vivo half-life orreduced dosage requirements) and hence may be preferred in somecircumstances. Isotopically labeled compounds of the invention cangenerally be prepared by following procedures analogous to thosedisclosed in the e.g., Examples herein by substituting an isotopicallylabeled reagent for a non-isotopically labeled reagent.

The term “prodrug” refers to compounds that are transformed in vivo toyield a disclosed compound or a pharmaceutically acceptable salt,hydrate or solvate of the compound. The transformation may occur byvarious mechanisms, such as through hydrolysis in blood. For example, ifa compound of the invention or a pharmaceutically acceptable salt,hydrate or solvate of the compound contains a carboxylic acid functionalgroup, a prodrug can comprise an ester formed by the replacement of thehydrogen atom of the acid group with a group such as (C₁-C₈)alkyl,(C₂-C₁₂)alkanoyloxymethyl, 1-(alkanoyloxy)ethyl having from 4 to 9carbon atoms, 1-methyl-1-(alkanoyloxy)-ethyl having from 5 to 10 carbonatoms, alkoxycarbonyloxymethyl having from 3 to 6 carbon atoms,1-(alkoxycarbonyloxy)ethyl having from 4 to 7 carbon atoms,1-methyl-1-(alkoxycarbonyloxy)ethyl having from 5 to 8 carbon atoms,N-(alkoxycarbonyl)aminomethyl having from 3 to 9 carbon atoms,1-(N-(alkoxycarbonyl)amino)ethyl having from 4 to 10 carbon atoms,3-phthalidyl, 4-crotonolactonyl, gamma-butyrolacton-4-yl,di-N,N-(C₁-C₂)alkylamino(C₂-C₃)alkyl (such as β-dimethylaminoethyl),carbamoyl-(C₁-C₂)alkyl, N,N-di(C₁-C₂)alkylcarbamoyl-(C₁-C₂)alkyl andpiperidino-, pyrrolidino- or morpholino(C₂-C₃)alkyl.

Similarly, if a compound of the invention contains an alcohol functionalgroup, a prodrug can be formed by the replacement of the hydrogen atomof the alcohol group with a group such as (C₁-C₆)alkanoyloxymethyl,1-((C₁-C₆)alkanoyloxy)ethyl, 1-methyl-1-((C₁-C₆)alkanoyloxy)ethyl(C₁-C₆)alkoxycarbonyloxymethyl, N-(C₁-C₆)alkoxycarbonylaminomethyl,succinoyl, (C₁-C₆)alkanoyl, α-amino(C₁-C₄)alkanoyl, arylacyl andα-aminoacyl, or α-aminoacyl-α-aminoacyl, where each α-aminoacyl group isindependently selected from the naturally occurring L-amino acids,P(O)(OH)₂, —P(O)(O(C₁-C₆)alkyl)₂ or glycosyl (the radical resulting fromthe removal of a hydroxyl group of the hemiacetal form of acarbohydrate).

If a compound of the invention incorporates an amine functional group, aprodrug can be formed by the replacement of a hydrogen atom in the aminegroup with a group such as R-carbonyl, RO-carbonyl, NRR′-carbonyl whereR and R′ are each independently (C₁-C₁₀)alkyl, (C₃-C₇)cycloalkyl,benzyl, or R-carbonyl is a natural a-aminoacyl or naturalα-aminoacyl-natural α-aminoacyl, —C(OH)C(O)OY¹ wherein Y¹ is H,(C₁-C₆)alkyl or benzyl, —C(OY²)Y³ wherein Y² is (C₁-C₄) alkyl and Y³ is(C₁-C₆)alkyl, carboxy(C₁-C₆)alkyl, amino(C₁-C₄)alkyl or mono-N— ordi-N,N—(C₁-C₆)alkylaminoalkyl, —C(Y⁴)Y⁵ wherein Y⁴ is H or methyl and Y⁵is mono-N— or di-N,N—(C₁-C₆)alkylamino, morpholino, piperidin-1-yl orpyrrolidin-1-yl.

The disclosure provides, at least in part, compounds represented byformula I, as depicted below. Also contemplated herein are compositionsthat include a compound represented by formula I and e.g., apharmaceutically acceptable carrier.

-   -   wherein X is C₁-C₃alkylene, optionally substituted with one, two        or three substituents selected from halogen or hydroxyl;    -   R₁ is selected from the group consisting of C₁-C₆alkyl,        C₃-C₆cycloalkyl, C₂-C₆alkenyl, and C₂-C₆alkynyl;    -   R₂ is selected from the group consisting of hydrogen and        C₁-C₆alkyl;    -   R₃ is independently selected, for each occurence from the group        consisting of hydrogen, C₁-C₆alkoxy, C₁-C₆alkyl, cyano,        C₃-C₆cycloalkyl, halogen, hydroxyl, and nitro;    -   R₄ is selected from the group consisting of hydrogen and        C₁-C₆alkyl;    -   R₅ is C₁-C₆alkyl;    -   or pharmaceutically acceptable salts or N-oxides thereof.

In one embodiment, R₁ can be C₁-C₆alkyl, such as methyl. In oneembodiment, R₂ can be hydrogen. In another embodiment, R₃ can beselected from the group consisting of hydrogen, C₁-C₆alkyl, halogen, andhydroxyl. In a further embodiment, R₃ can be hydrogen. In oneembodiment, R₄ and R₅ can each be C₁-C₆alkyl. In another embodiment, R₄may be hydrogen and R₅ may be methyl. In one embodiment, X may be(CH₂)_(n), wherein n is 1 or 2, such as 1.

In another embodiment, —NR₂—COR₁ can be in the meta position relative toX as shown in formula III.

In another embodiment, —NR₂—COR₁ can be in the para position relative toX as shown in formula IV.

The disclosure provides, at least in part, compounds represented byformula II, as depicted below. Also contemplated herein are compositionsthat include a compound represented by formula II and e.g., apharmaceutically acceptable carrier.

-   -   wherein R₁ is selected from the group consisting of C₁-C₆alkyl,        C₃-C₆cycloalkyl, C₂-C₆alkenyl, and C₂-C₆alkynyl;    -   R₂ is selected from the group consisting of hydrogen and        C₁-C₆alkyl;    -   R₃ is independently selected, for each occurence from the group        consisting of hydrogen, C₁-C₆alkoxy, C₁-C₆alkyl, cyano,        C₃-C₆cycloalkyl, halogen, hydroxyl, and nitro;    -   R₅ is hydrogen or C₁-C₆alkyl;        or pharmaceutically acceptable salts or N-oxides thereof.

Compounds of Formula V are also contemplated as shown below, as well ascompositions that include a compound represented by formula V and e.g.,a pharmaceutically acceptable carrier.

-   -   wherein R₁ is selected from the group consisting of C₁-C₆alkyl,        C₃-C₆cycloalkyl, C₂-C₆alkenyl, and C₂-C₆alkynyl;    -   R₃ is independently selected, for each occurence from the group        consisting of hydrogen, C₁-C₆alkoxy, C₁-C₆alkyl, cyano,        C₃-C₆cycloalkyl, halogen, hydroxyl, and nitro;    -   R₄ is selected from the group consisting of hydrogen and        C₁-C₆alkyl;    -   R₅ is hydrogen or C₁-C₆alkyl; and    -   A is a fused five or six membered heterocycle;        or pharmaceutically acceptable salts or N-oxides thereof.

In one embodiment, R₁ can be C₁-C₆alkyl, such as methyl. In anotherembodiment, R₁ and R₃ can each be C₁-C₆alkyl, such as methyl. In oneembodiment, R₂ can be hydrogen.

In some embodiments, a compound can be represented by

-   -   wherein p is 1 or 2;    -   R₁ is selected from the group consisting of C₁-C₆alkyl,        C₃-C₆cycloalkyl, C₂-C₆alkenyl, and C₂-C₆alkynyl;    -   R₄ and R₈ are each independently selected from the group        consisting of hydrogen and C₁-C₆alkyl;        or pharmaceutically acceptable salts or N-oxides thereof.

Contemplated compounds, and pharmaceutical compositions, comprising atleast one compound, may be selected from the group consisting of:N-acetyl-(R)-3-(4-aminophenyl)-2-methoxypropionic acid (Compound A),N-acetyl-(S)-3-(4-aminophenyl)-2-methoxypropionic acid (Compound B),racemic N-acetyl-3-(4-aminophenyl)-2-methoxypropionic acid (CompoundAB);

4-acetamino-N-hydroxy-2-methoxybenzamide;1-acetyl-6-methoxy-1,2,3,4-tetrahydroquinoline-5-carboxylic acid,5-acetamido-2hydroxybenzoic acid (e.g., acetalyated 5-aminosalicyclicacid) or pharmaceutically acceptable salts or N-oxides thereof.

The present disclosure also provides pharmaceutical compositionscomprising compounds as disclosed herein formulated together with one ormore pharmaceutically acceptable carriers. These formulations includethose suitable for oral, rectal, topical, buccal and parenteral (e.g.,subcutaneous, intramuscular, intradermal, or intravenous)administration, although the most suitable form of administration in anygiven case will depend on the degree and severity of the condition beingtreated and on the nature of the particular compound being used.

Therapeutic Applications

The disclosure further provides, in some embodiments, methods ofmodulating activity of one or more PPAR and/or EGF receptors comprisingexposing said receptor to a compound of the invention. For example,provided herein are methods of treating a disease associated withexpression or activity of one or more PPAR and/or EGF receptors in apatient comprising administering to the patient a therapeuticallyeffective amount of a compound of the invention.

One embodiment of the invention provides a method of treating tumors ofthe esophagus, stomach, pancreas, colon, prostate, breast, uterus,kidneys, and lungs comprising administering to a subject in need thereofa therapeutically effective amount of a compound of the invention. Alsocontemplated herein is a method for delaying clinical manifestation of acolorectal tumor, or a solid tumor (e.g., a breast, prostate, lung orhepatocellular carcinoma) in a patient, for example, a patient at riskof colorectal cancer, comprising administering to the patient aneffective amount of a compound disclosed herein. Administering such acompound may be on e.g., at least a daily basis. For example, the delayof clinical manifestation of a colorectal tumor in a patient as aconsequence of administering a compound disclosed here may be at leaste.g., 6 months, 1 year, 18 months or even 2 years or more as compared toa patient who is not administered a compound such as one disclosedherein.

Another embodiment of the invention provides a method of treating orameliorating chronic inflammation, such as Crohn's disease or ulcerativecolitis, comprising administering to a subject in need thereof atherapeutically effective amount of a compound of the invention.

Methods of treating dermatological conditions are also provided, such asthe treatment of at least one of: acne vulgaris, comedo-type acne,polymorphic acne, acne rosacea, nodulocystic acne, acne conglobata,senile acne, secondary acne, solar acne, acne medicamentosa oroccupational acne, ichthyosis, Darrier's disease, keratosis palmaris orplantaris, cutaneous, mucosal or ungual psoriasis, skin disorders due toexposure to UV radiation, of skin aging, photoinduced or chronologicalor actinic pigmentations and keratoses, acne hyperseborrhoea, simpleseborrhoea or seborrhoeic dermatitis, cicatrization disorders or stretchmarks. Methods of treating atopic dermatitis is also contemplated. Thecomposition may be administered orally or topically.

For example, continuous sebum production can increase in acne patients;and application of a sebum inhibitor, such as disclosed herein, may beuseful in the treatment of acne, seborrhea or alopecia. In anotherexample, chronic inflammation of hair follicles (keratinocytes) can bean indication of e.g., androgenic alopecia. An inhibitor of suchinflammation such as disclosed herein can be useful in e.g., thetreatment of hair loss.

Provided herein are methods of treating fine lines, wrinkles or surfaceirregularities of the skin, or protecting from and/or ameliorating freeradical damage to the skin, comprising topically administering aneffective amount of a composition comprising a compound of theinvention.

In some embodiments, a method of treating hair loss in a patientsuffering from unwanted hair loss is provided, comprising administeringan effective amount of a composition comprising a disclosed compound.Methods of treating alopecia areata, androgenetic alopecia and/ortelogenic defluvium are contemplated.

Also provided herein are methods of treating an age-related disorderselected from the group consisting of: diabetes, cataracts, Alzheimer'sdisease, Parkinson's disease, macular degeneration, retinal ulcers orretinal vasculitis, comprising administering an effective amount of acomposition comprising a disclosed compound. Also provided herein aremethods of treating a vascular or cardiac disorder, comprisingidentifying a patient suffering from or at risk of developing saiddisorder and administering to said patient an effective amount of adisclosed compound as defined above. For example, a cardiac disorderbeing treated may be chosen from chronic coronary ischemia,arteriosclerosis, congestive heart failure, ischemic or reperfusionrelated injury, angina, atherosclerosis, myocardial infarction, strokeand myocardial hypertrophy. In another embodiment, a method of treatingan autoimmune disorder is provided, wherein the autoimmune disorder maybe chosen from, for example, Addison's disease, chronic thyroiditis,dermatomyositis, Grave's disease, multiple sclerosis, systemic lupuserythematosis, psoriasis, or rheumatoid arthritis.

The compounds of the invention may be administered to patients (animalsand humans) in need of such treatment in dosages that will provideoptimal pharmaceutical efficacy. It will be appreciated that the doserequired for use in any particular application will vary from patient topatient, not only with the particular compound or composition selected,but also with the route of administration, the nature of the conditionbeing treated, the age and condition of the patient, concurrentmedication or special diets then being followed by the patient, andother factors which those skilled in the art will recognize, with theappropriate dosage ultimately being at the discretion of the attendantphysician. For treating clinical conditions and diseases noted above,the compound of this invention may be administered orally, topically,parenterally, by inhalation spray or rectally in dosage unitformulations containing conventional non-toxic pharmaceuticallyacceptable carriers, adjuvants and vehicles. The term parenteral as usedherein includes subcutaneous injections, intravenous, intramuscular,intrasternal injection or infusion techniques.

Generally, a therapeutically effective amount of active component willbe in the range of from about 0.1 mg/kg to about 100 mg/kg, optionallyfrom about 1 mg/kg to about 100 mg/kg, optionally from about 1 mg/kg to10 mg/kg. The amount administered will depend on variables such as thetype and extent of disease or indication to be treated, the overallhealth status of the particular patient, the relative biologicalefficacy of the compounds, formulation of compounds, the presence andtypes of excipients in the formulation, and the route of administration.The initial dosage administered may be increased beyond the upper levelin order to rapidly achieve the desired blood-level or tissue level, orthe initial dosage may be smaller than the optimum and the daily dosagemay be progressively increased during the course of treatment dependingon the particular situation. Human dosage can be optimized, e.g., in aconventional Phase I dose escalation study designed to run from 0.5mg/kg to 20 mg/kg. Dosing frequency can vary, depending on factors suchas route of administration, dosage amount and the disease conditionbeing treated. Exemplary dosing frequencies are once per day, once perweek and once every two weeks.

Contemplated formulations or compositions comprise a disclosed compoundand typically include a compound a pharmaceutically acceptable carrier.

Contemplated compositions may be administered by various means,depending on their intended use, as is well known in the art. Forexample, if compositions of the present invention are to be administeredorally, they may be formulated as tablets, capsules, granules, powdersor syrups. Alternatively, formulations of the present invention may beadministered parenterally as injections (intravenous, intramuscular orsubcutaneous), drop infusion preparations or enemas or suppositories.For application by the ophthalmic mucous membrane route, compositions ofthe present invention may be formulated as eyedrops or eye ointments.These formulations may be prepared by conventional means, and, ifdesired, the compositions may be mixed with any conventional additive,such as an excipient, a binder, a disintegrating agent, a lubricant, acorrigent, a solubilizing agent, a suspension aid, an emulsifying agentor a coating agent.

In formulations of the subject invention, wetting agents, emulsifiersand lubricants, such as sodium lauryl sulfate and magnesium stearate, aswell as coloring agents, release agents, coating agents, sweetening,flavoring and perfuming agents, preservatives and antioxidants may bepresent in the formulated agents.

Subject compositions may be suitable for oral, nasal, topical (includingbuccal and sublingual), rectal, vaginal, aerosol and/or parenteraladministration. The formulations may conveniently be presented in unitdosage form and may be prepared by any methods well known in the art ofpharmacy. The amount of composition that may be combined with a carriermaterial to produce a single dose vary depending upon the subject beingtreated, and the particular mode of administration.

Methods of preparing these formulations include the step of bringinginto association compositions of the present invention with the carrierand, optionally, one or more accessory ingredients. In general, theformulations are prepared by uniformly and intimately bringing intoassociation agents with liquid carriers, or finely divided solidcarriers, or both, and then, if necessary, shaping the product.

Formulations suitable for oral administration may be in the form ofcapsules, cachets, pills, tablets, lozenges (using a flavored basis,usually sucrose and acacia or tragacanth), powders, granules, or as asolution or a suspension in an aqueous or non-aqueous liquid, or as anoil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup,or as pastilles (using an inert base, such as gelatin and glycerin, orsucrose and acacia), each containing a predetermined amount of a subjectcomposition thereof as an active ingredient. Compositions of the presentinvention may also be administered as a bolus, electuary, or paste.

In solid dosage forms for oral administration (capsules, tablets, pills,film-coated tablets, sugar-coated tablets, powders, granules and thelike), the subject composition is mixed with one or morepharmaceutically acceptable carriers, such as sodium citrate ordicalcium phosphate, and/or any of the following: (1) fillers orextenders, such as starches, lactose, sucrose, glucose, mannitol, and/orsilicic acid; (2) binders, such as, for example, carboxymethylcellulose,alginates, gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; (3)humectants, such as glycerol; (4) disintegrating agents, such asagar-agar, calcium carbonate, potato or tapioca starch, alginic acid,certain silicates, and sodium carbonate; (5) solution retarding agents,such as paraffin; (6) absorption accelerators, such as quaternaryammonium compounds; (7) wetting agents, such as, for example, acetylalcohol and glycerol monostearate; (8) absorbents, such as kaolin andbentonite clay; (9) lubricants, such a talc, calcium stearate, magnesiumstearate, solid polyethylene glycols, sodium lauryl sulfate, andmixtures thereof; and (10) coloring agents. In the case of capsules,tablets and pills, the compositions may also comprise buffering agents.Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugars, as well as high molecular weight polyethylene glycolsand the like.

Formulations and compositions may include micronized crystals of thedisclosed compounds. Micronization may be performed on crystals of thecompounds alone, or on a mixture of crystals and a part or whole ofpharmaceutical excipients or carriers. Mean particle size of micronizedcrystals of a disclosed compound may be for example about 5 to about 200microns, or about 10 to about 110 microns.

A tablet may be made by compression or molding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared usingbinder (for example, gelatin or hydroxypropylmethyl cellulose),lubricant, inert diluent, preservative, disintegrant (for example,sodium starch glycolate or cross-linked sodium carboxymethyl cellulose),surface-active or dispersing agent. Molded tablets may be made bymolding in a suitable machine a mixture of the subject compositionmoistened with an inert liquid diluent. Tablets, and other solid dosageforms, such as film coated tablets or sugar coated tablets, capsules,pills and granules, may optionally be scored or prepared with coatingsand shells, such as enteric coatings and other coatings well known inthe pharmaceutical-formulating art.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups andelixirs. In addition to the subject composition, the liquid dosage formsmay contain inert diluents commonly used in the art, such as, forexample, water or other solvents, solubilizing agents and emulsifiers,such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, oils (in particular, cottonseed, groundnut, corn, germ, olive,castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethyleneglycols and fatty acid esters of sorbitan, cyclodextrins and mixturesthereof.

Suspensions, in addition to the subject composition, may containsuspending agents as, for example, ethoxylated isostearyl alcohols,polyoxyethylene sorbitol and sorbitan esters, microcrystallinecellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth,and mixtures thereof.

Formulations for rectal or vaginal administration may be presented as asuppository, which may be prepared by mixing a subject composition withone or more suitable non-irritating excipients or carriers comprising,for example, cocoa butter, polyethylene glycol, a suppository wax or asalicylate, and which is solid at room temperature, but liquid at bodytemperature and, therefore, will melt in the body cavity and release theactive agent. Formulations which are suitable for vaginal administrationalso include pessaries, tampons, creams, gels, pastes, foams or sprayformulations containing such carriers as are known in the art to beappropriate.

Dosage forms for transdermal or topical administration of a subjectcomposition include powders, sprays, ointments, pastes, creams, lotions,gels, solutions, patches and inhalants. The active component may bemixed under sterile conditions with a pharmaceutically acceptablecarrier, and with any preservatives, buffers, or propellants which maybe required.

The ointments, pastes, creams and gels may contain, in addition to asubject composition, excipients, such as animal and vegetable fats,oils, waxes, paraffins, starch, tragacanth, cellulose derivatives,polyethylene glycols, silicones, bentonites, silicic acid, talc and zincoxide, or mixtures thereof.

Powders and sprays may contain, in addition to a subject composition,excipients such as lactose, talc, silicic acid, aluminum hydroxide,calcium silicates and polyamide powder, or mixtures of these substances.Sprays may additionally contain customary propellants, such aschlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, suchas butane and propane.

Compositions and compounds of the present invention may alternatively beadministered by aerosol. This is accomplished by preparing an aqueousaerosol, liposomal preparation or solid particles containing thecompound. A non-aqueous (e.g., fluorocarbon propellant) suspension couldbe used. Sonic nebulizers may be used because they minimize exposing theagent to shear, which may result in degradation of the compoundscontained in the subject compositions.

Ordinarily, an aqueous aerosol is made by formulating an aqueoussolution or suspension of a subject composition together withconventional pharmaceutically acceptable carriers and stabilizers. Thecarriers and stabilizers vary with the requirements of the particularsubject composition, but typically include non-ionic surfactants(Tweens, Pluronics, or polyethylene glycol), innocuous proteins likeserum albumin, sorbitan esters, oleic acid, lecithin, amino acids suchas glycine, buffers, salts, sugars or sugar alcohols. Aerosols generallyare prepared from isotonic solutions.

Pharmaceutical compositions of this invention suitable for parenteraladministration comprise a subject composition in combination with one ormore pharmaceutically-acceptable sterile isotonic aqueous or non-aqueoussolutions, dispersions, suspensions or emulsions, or sterile powderswhich may be reconstituted into sterile injectable solutions ordispersions just prior to use, which may contain antioxidants, buffers,bacteriostats, solutes which render the formulation isotonic with theblood of the intended recipient or suspending or thickening agents.

Examples of suitable aqueous and non-aqueous carriers which may beemployed in the pharmaceutical compositions of the invention includewater, ethanol, polyols (such as glycerol, propylene glycol,polyethylene glycol, and the like), and suitable mixtures thereof,vegetable oils, such as olive oil, and injectable organic esters, suchas ethyl oleate and cyclodextrins. Proper fluidity may be maintained,for example, by the use of coating materials, such as lecithin, by themaintenance of the required particle size in the case of dispersions,and by the use of surfactants. The efficacy of treatment with thesubject compositions may be determined in a number of fashions known tothose of skill in the art.

Throughout the description, where compositions are described as having,including, or comprising specific components, it is contemplated thatcompositions also consist essentially of, or consist of, the recitedcomponents. Similarly, where processes are described as having,including, or comprising specific process steps, the processes alsoconsist essentially of, or consist of, the recited processing steps.Except where indicated otherwise, the order of steps or order forperforming certain actions are immaterial so long as the inventionremains operable. Moreover, unless otherwise noted, two or more steps oractions may be conducted simultaneously.

EXAMPLES

The compounds of the present invention can be prepared in a number ofways well known to one skilled in the art of organic synthesis. Morespecifically, compounds of the invention may be prepared using thereactions and techniques described herein. In the description of thesynthetic methods described below, it is to be understood that allproposed reaction conditions, including choice of solvent, reactionatmosphere, reaction temperature, duration of the experiment and workupprocedures, can be chosen to be the conditions standard for thatreaction, unless otherwise indicated. It is understood by one skilled inthe art of organic synthesis that the functionality present on variousportions of the molecule should be compatible with the reagents andreactions proposed. Substituents not compatible with the reactionconditions will be apparent to one skilled in the art, and alternatemethods are therefore indicated. The starting materials for the examplesare either commercially available or are readily prepared by standardmethods from known materials.

Example 1 Preparation ofN-acetyl-(R)-(−)-3-(4-aminophenyl)-2-methoxypropionic acid (N-AcetylE2): Compound A

To (R)-(−)-3-(4-aminophenyl)-2-methoxypropionic acid (40 g) in a 0.5 Lglass reactor was added ethyl acetate (80 g) and acetic anhydride (62.8g). The mixture was stirred at 90° C. for 1 hour. Upon cooling, thesolvent was removed by vaccum distillation, providing an oily residue.To this residue was added water (120 g) and ethyl acetate (120 g). Afterstirring for 10 min at 35° C., the layers were separated and the aqueouslayer discarded. The organic layer solvent was removed by vacuumdistillation. Acetone (120 g) was then added and the resulting mixturewas warmed until dissolution was complete. The solution was cooled to 0°C., and the product precipitated which was collected by filtration. Thesolid was rinsed with acetone (20 g) and dried at 65° C. to afford 26 gof the title compound.

Example 2 Docking Studies

The binding of compounds A, B, and their non-acetylated derivatives, (aswell as 5-aminosalicyclic acid (5-ASA) and 5-acetamido-hydroxybenzoicacid) to PPARγ and PPARα receptors is evaluated.

While 5-ASA shows good affinity for PPARγ, the N-acetylation of 5-ASAled to a rigid linear structure that did not occupy the active site in aoptimal way. A loss of the hydrogen bond between H449 and the phenolicgroup of the compound occurred which may explain the inactivity ofNAc-5-ASA. The binding of compound B and its non-acetylated counterpartinto PPARγ indicated that these compounds may activate the receptorbased on the receptor's binding structural prerequisites. Superpositionof the compounds indicated that they bind to different parts of theactive site.

The binding of compound A and its non acetylated counterpart3-(4-aminophenyl)-2-methoxypropanoic acid) into PPARγ indicated thatthese compounds may also activate the receptor. In contrast to compoundB, the superposition of compound A free amine and N-acetyl derivativeshow that they occupy the same portion of the active site, indicating apossible similarity in activity.

Example 3 Anti-Murine Colitis Model

Colitis in C57bI6 mice is induced by administering TNBS (150 mg/kg) byoral gavage on day −3. Stool samples were taken 8 hours afteradminstration. At day 0-5, N-acetylated E2 (30mM) was administrated byoral gavage. On day 5, micewere analyzed to obtain a mortalitymacroscopic score (Wallace, Gastroenterology 96: 29-36, 1989).

As shown in FIG. 1, mortality was comparable between mice given TNBSversus those given TNBS and N-acetyl E2. However, as shown in FIG. 2,the level of colonic lesions was significantly less in mice given TNBSand N-acetyl E2 compared to those given TNBS alone. FIG. 3 demonstrateshow MPO (myleoperoxidase) decreased with administration of N-acetyl E2with TNBS. In FIG. 4, the Ameho score (Gut 41: 487-493, 1997) indicatedthat colonic inflammation had decreased with adminstration of N-acetylE2 with TNBS.

Example 4 Keratinocytes

To assess the possible toxic or cytostatic effect of the substancesunder study, a spectrophotometric test (MTT) was carried out. The humanprimary keratinocytes, isolated from skin biopsies, were plated in wellsof a 24-well plate in suitable medium with addition of antibiotics,calcium, and specific growth factors. At around 70% confluence, thecells were exposed to the presence of Compound A, at variousconcentrations (0.1-1-2 mM), for 24 and 48 h in suitable medium withaddition of antibiotics, calcium, but no growth factors. This culturecondition was done for all the subsequent experiments. At the end of thetreatment, the MTT test was done. The results are indicated in FIG. 5.Compound A in all concentrations used did not show any effect oncellular vitality.

Example 5 TNF Alpha

Analysis of the inhibition of compound Aof the mRNA induction of theproinflammatory cytokine TNF-alpha by H₂O₂ was carried out by Real timeRT-PCR. The keratinocytes were plated in dishes of 6 cm/Ø. At 80%confluence, the cells were treated with H₂O₂ (300 μM) in presence ofCompound A at the three concentrations (0.01-0.1-0.5 mM) for 6 h. At theend of the treatment, the cells were lysed in a lysis buffer andsubjected to isolation and subsequent retrotranscription of the RNA.Compound A proved able to inhibit the expression of the mRNA of TNF-αinduced by H₂O₂ at the two higher doses (0.1 mM; 0.5 mM). The higherdose demonstrated a complete inhibition of the proinflammatory cytokinewith an effect similar to troglitazone (Tg). (FIG. 6)

Example 6 Inhibition of mRNA Expression of IL-6 Induced by Presence ofIFN-γ

Analysis of the inhibition by compound A of the mRNA induction of theproinflammatory cytokine IL-6 by IFN-γ was done through Real timeRT-PCR. The keratinocytes were plated in dishes of 6 cm/Ø.

At 80% confluence, the cells were treated with IFN-γ (30 ng/ml) inpresence of compound A (N-Acetylged) at the three concentrations(0.01-0.1-0.5 mM) for 6 h. At the end of the treatment, the cells werelysed in a lysis buffer and subjected to isolation and subsequentretrotranscription of the RNA The results (as shown in FIG. 7) revealthe ability of Compound A to inhibit the expression of the inflammatorycytokine induced by presence of IFN-γ which does not appear to bedose-dependent.

Example 7 Inhibitory Capacity on the Activation of Nuclear Factor NF-kBInduced by Presence of H₂O₂

Evaluation of the inhibition by compound A of the activation of nucleartranscription factor NF-kB induced by the presence of H₂O₂ was done byanalysis in cytofluorimetry.

The keratinocytes were plated in wells of a 12-well plate. At 80%confluence, the cells were treated with H₂O₂ (300 μM) in presence ofcompound A at the three concentrations (0.01-0.1-0.5 mM) for 1 h. At theend of the treatment, the cells were fixed in paraformaldehyde,permeabilized in methanol and then incubated in presence of the specificantibody of subunit p65. Compound A revealed an inhibitory effect on theactivation and subsequent translocation of NF-kB in dose-dependentmanner (FIG. 8).

Example 8 Inhibition of Protein Expression of IL-6 Induced by Presenceof LPS

Analysis of the inhibition by Compound A of the protein induction ofIL-6 by LPS (lipopolysaccharide) was done with the ELISA kit. Thekeratinocytes were plated in wells of a 24-well plate. At 80%confluence, the cells were treated with LPS (10 μg/ml) in presence ofcompound A at the three concentrations (0.01-0.1-0.5 mM) for 24 h. Atthe end of the treatment, the supernatant was decanted, centrifuged soas to remove any cell detritus, and kept at −80° C. until the time ofthe analysis. The quantity of IL-6 present in the supernatant wasnormalized by the protein concentration of the sample itself. Theresults (FIG. 9) revealed the ability of compound A to inhibit, indose-dependent manner, the protein expression of the inflammatorycytokine under study.

Example 9—Human Sebocytes

To assess the possible toxic or cytostatic effect of the substancesunder study, a spectrophotometric test (MTT) was carried out. Thesebocytes were plated in wells of a 24-well plate in suitable mediumwith addition of antibiotics, calcium and EGF. At roughly 70%confluence, the cells were exposed to the presence of compound A, invarious concentrations (0.1-0.5-1-2 mM), for 24 and 48 h. At the end ofthe treatment, the MTT test was performed. Compound A in allconcentrations used demonstrated no effects on cell vitality. (FIG. 10)

Example 10 Evaluation of the Inhibitory Capacity of a Compound onSebogenesis Induced by Stimuli of Lipid Type (Linoleic Acid,Testosterone)

Analysis of the inhibition by (compound A) of sebogenesis induced bytreatment with linoleic acid (LA) and with testosterone (TST) wasevaluated by spectrofluorimetry, using Nile Red as selective marker ofintracellular lipids (Nile Red Assay). The sebocytes were plated inwells of a 24-well plate. Next day, they were deprived of serum (2%) andafter 24 h they were stimulated, for another 24 h, with LA (10-4M), TST(20 nM) in presence or in absence of A (1 mM). At the end of thetreatment, the sebocytes were stained with Nile Red. The quantitativeanalysis was done by spectrofluorimetry, which made it possible todistinguish between neutral lipids and polar lipids based on thedifferent wavelength of excitation and emission. The data obtainedrevealed that the treatment with LA is able to induce lipid synthesisand that the combined LA+TST treatment further increases this effect.The presence of Compound A proved able to reduce the lipidogenicstimulus. (FIG. 11).

Example 11 Evaluation of the Inhibitory Capacity on Sebogenesis Inducedby Stimuli of Lipid Type (Linoleic Acid, Testosterone): Evaluation ofFatty Acids and Squalene

In order to evaluate in greater detail the inhibition by Compound A ofsebogenesis induced by LA and TST, assays were performed on the lipidextract of the sebocytes using gas chromatography coupled with massspectrometry (GC-MS). The sebocytes were treated by the scheme describedfor the Nile Red assay. At the end of the treatment, the cells wereremoved and then the lipid extraction was done by using organicsolvents. One part of the extract was used to analyze the fatty acidcomposition, while the other part was used for the determination of thequantity of squalene, a lipid characteristic of sebum. The fatty acidassay showed that the lipidogenic stimulus induced by the treatment withLA and LA+TST was reduced by the presence of A (FIG. 12A). These resultsare confirmed by the squalene analysis. (FIG. 12B)

Example 12 Evaluation of the Inhibitory Capacity on Sebogenesis Inducedby Stimuli of Lipid Type (Linoleic Acid, Testosterone)

Analysis of the inhibition by compound A of sebogenesis induced bytreatment with linoleic acid (LA) and with testosterone (TST) wasevaluated by spectrofluorimetry, using Nile Red as selective marker ofintracellular lipids (Nile Red Assay). The sebocytes were plated inwells of a 24-well plate. Next day, they were deprived of serum (2%) andafter 24 h they were stimulated, for another 24 h, with LA (10-4M), TST(20 nM) in presence or in absence of compound A (1 mM). At the end ofthe treatment, the sebocytes were stained with Nile Red. Thequantitative analysis was done by spectrofluorimetry, which made itpossible to distinguish between neutral lipids and polar lipids based onthe different wavelength of excitation and emission. The data obtained(FIG. 13) revealed that the treatment with LA is able to induce lipidsynthesis and that the combined LA+TST treatment further increases thiseffect. The presence of A proved able to reduce the lipidogenicstimulus. No differences were observed in regard to the times oftreatment with the A.

References

All publications and patents mentioned herein, including those itemslisted below, are hereby incorporated by reference in their entirety asif each individual publication or patent was specifically andindividually incorporated by reference. In case of conflict, the presentapplication, including any definitions herein, will control.

Equivalents

While specific embodiments of the subject invention have been discussed,the above specification is illustrative and not restrictive. Manyvariations of the invention will become apparent to those skilled in theart upon review of this specification. The full scope of the inventionshould be determined by reference to the claims, along with their fullscope of equivalents, and the specification, along with such variations.

Unless otherwise indicated, all numbers expressing quantities ofingredients, reaction conditions, and so forth used in the specificationand claims are to be understood as being modified in all instances bythe term “about.” Accordingly, unless indicated to the contrary, thenumerical parameters set forth in this specification and attached claimsare approximations that may vary depending upon the desired propertiessought to be obtained by the present invention.

1.-19. (canceled)
 20. The compoundN-acetyl-(S)-3-(4-aminophenyl)-2-methoxypropionic acid, or apharmaceutically acceptable salt or an N-oxide thereof.
 21. Apharmaceutical composition comprising the compound of claim 20, or apharmaceutically acceptable salt or an N-oxide thereof; and apharmaceutically acceptable carrier.
 22. The compoundN-acetyl-(S)-3-(4-aminophenyl)-2-methoxypropionic acid, or apharmaceutically acceptable salt thereof.
 23. A pharmaceuticalcomposition comprising the compound of claim 22, or a pharmaceuticallyacceptable salt thereof and a pharmaceutically acceptable carrier. 24.The compound N-acetyl-(S)-3-(4-aminophenyl)-2-methoxypropionic acid. 25.A pharmaceutical composition comprising the compound of claim 24; and apharmaceutically acceptable carrier.